Development and evaluation of an immunoassay for the monitoring of the anti-HIV drug amprenavir

Erica Bastiani, Fabio Benedetti, Federico Berti, Pietro Campaner, Elena Donadel, Michela Montagna, Mario Regazzi, Serena Rinaldi, Adriano Savoini, Romina Venturini

Research output: Contribution to journalArticle

Abstract

An assay for routine therapeutic drug monitoring of anti-HIV HAART drugs in clinical use is highly desirable, in order to rapidly measure the pharmacokinetic parameters on single patients. We have started a project to develop a panel of enzyme-linked immunosorbent assays (ELISA) for the whole set of HAART drugs, and the development, performance and evaluation of the assay for amprenavir is described here. A diazo conjugate of amprenavir has been used in order to raise polyclonal anti-amprenavir antibodies in rabbits. Antisera have been used to set up a quantitative and rapid competitive assay. Plasma samples are simply diluted in the assay buffer after thermal inactivation, before running the assay. The assay allows the detection of amprenavir in the quantification range 400-5000 ng/ml, in a diluted plasma sample. The assay has been compared with an HPLC reference technique, on 27 samples from treated patients. Within the quantification range, the ELISA data are well correlated with the HPLC results by a regression line close to the identity, and a Bland-Altman analysis shows the agreement between the two methods.

Original languageEnglish
Pages (from-to)35-41
Number of pages7
JournalJournal of Immunological Methods
Volume325
Issue number1-2
DOIs
Publication statusPublished - Aug 31 2007

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Keywords

  • Drug monitoring
  • ELISA
  • HAART therapy
  • HIV

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

Cite this

Bastiani, E., Benedetti, F., Berti, F., Campaner, P., Donadel, E., Montagna, M., Regazzi, M., Rinaldi, S., Savoini, A., & Venturini, R. (2007). Development and evaluation of an immunoassay for the monitoring of the anti-HIV drug amprenavir. Journal of Immunological Methods, 325(1-2), 35-41. https://doi.org/10.1016/j.jim.2007.05.009